Method for recovering and purifying trophoblastic beta-1-glycoprotein
SUBSTANCE: method for recovering and purifying trophoblastic beta-1-glycoprotein (TBG) from retroplacental blood serum by ammonium sulphate fractionation, washing and centrifugation of the solution residual matter, affinity sepharose chromatography followed by elution and lyophilisation of an end product. The retroplacental blood is centrifugated at 10000 rpm within one hour. Sodium chloride is added to a supernatant to end concentration 0.1-1.0 mol/l, while triton X-100 is added to end concentration 0.01-0.9%. Centrifugation is performed once again. The prepared serum is saturated with ammonium sulphate to 30%, incubated within a night in stirring, centrifugated. The deposit is threefold washed in 30% ammonium sulphate, suspended in 0.15M phosphate buffered saline pH 7.0-7.5, containing 0.5 M sodium chloride and 0.05% triton X-100, dialysed in an ultrafiltration cell, with tenfold amount of the buffered saline passed, five-fold diluted and ultrafiltered at filtration limit 100000 D. The prepared permeate is processed with single affinity sepharose chromatography with immobilised monoclonal TBG antibodies. The affinity-bound TBG is eluted with 0.1M glycine-Hcl buffer solution pH 2.55. The eluate is immediately neutralised, concentrated, dialysed against physiological saline and lyophilised.
EFFECT: higher yield and purity of the end product.
2 ex, 1 tbl
The invention relates to pharmaceutical industry and can be used in medicine, biotechnology for producing drug USSR β-1-glycoprotein (TBG), used as a component of diagnostic systems and tools for the treatment of cancer, autoimmune and allergic diseases. The method allows to increase the yield and purity of the target product.
It is known that TBG, as a specific marker of pregnancy, is synthesized by the syncytiotrophoblast cells and secreted into the bloodstream of the mother. According to most authors TBG is a glycoprotein containing up to 28% carbohydrates and having a molecular weight of 90000 Yes (Bohn N., 1974) to 113000 Yes (Tatarinov US and others, 1974). However, in the literature there are data that TBG forms a family consisting of 4 proteins with a molecular mass of 72, 64, 62 and 54 kDa (Plouzek S.A., J.Y. Chou, 1991). Other authors believe that TBG is a heterogeneous protein heterogeneity due to the presence of two options TBG - alpha and beta (Pola A. et al., 1992). When this molecular weight TBG-alpha equal to 110 KD, and TBG-beta 75 KD (M. Ito et al., 1981). The absence of well-attested data about the molecular heterogeneity of TBG and accurate description of the structure of the molecule is largely the reason for the lack of an effective system of selection and ochistitel to obtain TBG is retroplatsentarno blood.
Known multi-step allocation method of TBG retroplatsentarno human blood . Serum obtained from retroplatsentarno blood was fractionally consistently with rivanol and ammonium sulfate, and then subjected to gel filtration on Sephadex G-25. The resulting material was subjected to ion exchange chromatography on DEAE-cellulose. The fractions containing TBG, United, fractionally ammonium sulfate and were gel-filtered on Sephadex G-200. After another alpacamania fractionation and dialysis was performed chromatography on a column of hydroxylapatite and after clarification by centrifugation, ion-exchange chromatography on microgranular cellulose KM-32. The final stage of purification was the preparative isoelectrofocusing on impaling with a range of 3.0 to 10.0. The result of the entire purification procedure was to obtain the drug TBG with purification efficiency of 93%. The yield was 1.1%.
The cleaning procedure allowed to obtain 1.8 mg relatively clean of the desired protein from a sample containing 160 mg, involves two procedures gel-filtration, two ion-exchange, one hydrophobic chromatography, three procedures alpacamania fractionation, isoelectrofocusing.
Thus, the apparent disadvantages of the described method are extraordinary complexity and trudem is here, very high cost, especially considering the extremely low yield of the target product, equal to 1.1%.
There is a method of allocation of TBG waste of gammaglobulin of retroplatsentarno blood , providing for the suspension of a solid residue containing TBG in water, centrifugation and subsequent alpacamania fractionation. After 4 days of dialysis and centrifugation of the resulting supernatant is treated with hydroxyapatite and Proskurov 30-60 minutes, and again centrifuged. The supernatant is subjected to concentration ultrafilter and the two-day dialysis, then centrifuged and lyophilizers. Dried product is dissolved in phosphate buffer and subjected to column chromatography on hydroxyapatite. The eluate cialiswhat within 36 hours and lyophilizers. The output is 35.5%, purity of the drug - 95%. The described method is superior to previous and purity of the drug, and exit, but also very time-consuming, takes significant time and leads to unnecessary losses due to the numerous procedures of dialysis.
The same problems burdened methods proposed Hao et al.  and Wasiliana et al. that when the purity of the target product (95%) are accompanied by significantly lower yields (20%).
There is a method of extracting the Oia TBG serum retroplatsentarno blood or sediment, obtained in the production of gamma globulin from the serum retroplatsentarno blood , providing alpacamania fractionation followed by dialysis for 4 days. After centrifugation the supernatant is subjected to ion-exchange chromatography. The eluate concentrate and cialiswhat within 72 hours. Otvetsvennyy the preparation is centrifuged and passed through the sorbent, containing lectin (concanavalin a). Elyuirovaniya fraction containing TBG, dialist and lyophilizers. The release of the drug in the purity of 96% is 20%.
In the so-called BIC options described ion-exchange chromatography on hexylsilane laid high quantitative loss of protein. The duration of the method, and the low yield of the target product does not allow to consider the described method as a solution to the problem of isolation and purification of this protein complex as USSR beta-1-glycoprotein.
The closest analogue may be considered the method proposed Wageline et al. . The authors used as source material retroplatsentarno blood or residue obtained in the production of gamma globulin. To reduce procedure time, the authors received alpacamania residue, which was subjected sequentially gel-filtration chromatography on Sephadex is -100, ion-exchange chromatography on DEAE-cellulose and chromatography on sepharose with immobilized zibarro-blue dye. At the end of the process, the resulting product was again subjected to gel filtration on Sephadex G-200. The authors rash called chromatography on zibarro-sepharose affine, while is hydrophobic procedure. The multiple steps involved in the extraction procedure determines its complexity and low yield of the target product (about 25%) with good levels of purity (not less than 96%).
An object of the invention is to increase the yield and purity allocated TBG. This object is achieved as follows.
Proven in the absence of hepatitis b virus, antibodies to HIV, hepatitis C retroplatsentarno blood is centrifuged, add sodium chloride (0.1 to 1.0 mol/l) and Triton X-100 (0.01 and 0.1%) for blocking of nonspecific adsorption and inactivation of viruses, and again centrifuged. The thus prepared serum saturated with ammonium sulfate to 30%, incubated over night at +4°C and stirring and centrifuged. The precipitate washed three times 30% ammonium sulfate, suspended in 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride, and cialiswhat in the ultrafiltration cell, missing a 10-fold volume of buffer. The resulting diluted 5 times with 0.1 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride and 0.05% Triton X-100, and subjected to ultrafiltration in the ultrafiltration cell with a filter to limit exceptions 100000 Yes. The resulting permit put on affinity sorbent with immobilized monoclonal antibodies to TBG. Nonspecific sorbiruyushchiesya components of the serum wash with sorbent consistently 4-5 volumes of 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride, and the same volume of 0.15 M sodium chloride solution. Affine-associated TBG elute with 0.1 M glycine-HCl buffer solution.
The eluate quickly neutralized to a pH of 6.5-7.5, dialist against saline and lyophilizers.
Crucial advantage of the proposed method is able to obtain the drug TBG with a high degree of purity and effective yield of the target product.
The method is illustrated by the following examples.
Example 1. All procedures performed at +4°C. 200 ml retroplatsentarno blood centrifuged with a speed of 10,000 rpm for 1 hour, the precipitate discarded, and the supernatant add sodium chloride to a final concentration of 0.5 M and Triton X-100 to a final concentration of 0.05%. The centrifugation repeated. Serum containing 31 mg TBG, thus prepared, is saturated with ammonium sulfate to 30% saturation, incubated overnight with stirring and cent is fugeret with a speed of 10,000 rpm for 1 hour. The precipitate washed three times 30% ammonium sulfate, suspended in 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride and 0.05% Triton X-100, dialist in the ultrafiltration cell, missing a 10-fold volume of buffer. To the drug add 4 volumes of 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride and 0.05% Triton X-100 and subjected to ultrafiltration in the ultrafiltration cell using a filter with a limit exceptions 100000 Yes to complete the passage of the solution through the filter. The resulting permeate is combined with 40 ml sepharose CL-6B, with "sewn" monoclonal antibodies to TBG. Incubation is carried out for 12 h with constant stirring on an orbital shaker.
Nonspecific sorbiruyushchiesya components of the serum wash with sorbent consistently 4-5 volumes of phosphate-saline buffer solution containing 0.5 M sodium chloride solution and the same amount of 0.15 M solution of sodium chloride. The sorbent is placed in a chromatographic column of 2.5×8,5 see After washing the sorbent solution of sodium chloride conduct elution with 0.1 M glycine-Hcl buffer solution with a pH of 2.55 at 80 ml/h, while in optical density at a wavelength of 280 nm. The elution is carried out to achieve zero values of optical density. The fraction containing the target product immediately neutralized to a pH of 6.5-7.5, concentrate, dialist in ultrafiltration cell relative to saline and lyophilizers.
The purity of the obtained product, analyzed by electrophoresis in a density gradient polyacrylamide gel, is 98%. The output of the preparation according to the immunoassay of 23.3 mg (75%). The drug test for the absence of antibodies to HIV, hepatitis b virus and antibodies to hepatitis C.
Example 2. All procedures performed at +4°C. 200 ml retroplatsentarno blood centrifuged with a speed of 10,000 rpm for 1 hour, the precipitate discarded, and the supernatant add sodium chloride to a final concentration of 0.5 M and Triton X-100 to a final concentration of 0.05%. The centrifugation repeated. Serum containing 31 mg TBG, thus prepared, is saturated with ammonium sulfate to 30% saturation, incubated overnight with stirring and centrifuged with a speed of 10,000 rpm for 1 hour. The precipitate washed three times 30% ammonium sulfate, suspended in 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride and 0.05% Triton X-100, dialist in the ultrafiltration cell, missing a 10-fold volume of buffer. To the drug add 4 volumes of 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride and 0.05% Triton X-100, and all Laut ultrafiltration in the ultrafiltration cell using a filter with a limit exceptions 100000 Yes to complete the passage of the solution through the filter. The permeate is applied to the chromatographic column of 2.5×8,5 filled with 40 ml sepharose CL-6B, with "sewn" monoclonal antibodies to TBG, equilibriating phosphate-saline buffer solution. The rate of deposition of 40 ml/hour, a Drawing exercise three times the passage of the applied volume through the column.
Nonspecific sorbiruyushchiesya components of blood wash with sorbent consistently phosphate-saline buffer solution containing 0.5 M sodium chloride, and 0.15 M sodium chloride at a rate of 80 ml/h until reaching zero values of optical density.
After washing the sorbent is carried out the elution of 0.1 M glycine-HCl buffer solution with a pH of 2.55 and a rate of 40 ml/hour elution Process control by optical density at a wavelength of 280 nm. Elution is carried out until reaching zero values of optical density. The fraction containing the target product, immediately neutralized to a pH of 6.5-7.5, concentrate, dialist in ultrafiltration cell relative to saline and lyophilizers.
The purity of the obtained product, analyzed by electrophoresis in a density gradient polyacrylamide gel, is 98%. The output of the preparation according to the immunoassay is 25.1 mg (81%). The drug test for the absence of antibodies to HIV, hepatitis b virus and antibodies to the virus of hepatitis is itâ C.
Using the proposed method allows to obtain the target product with a high purity (at least 98%) and high: 75-81% yield. Step-by-step description of procedures for the isolation and purification are shown in table 1.
|Example 1||Example 2|
|No.||Operation||Volume of sample (ml)||Qty TBG (mg)||Volume of sample (ml)||Qty TBG (mg)|
Sources of information
1. Aveskulov, Haasse, Nvenergy, Justiniano. Isolation and purification of specific β1-g-globulin. "Questions of medical chemistry, 1978, No. 24, s-244.
2. Swimers, Schrivener, Appalling, Usepagov, Justiniano. The method of obtaining the USSR beta-1-glycoprotein from waste retroplatsentarno blood in the production of gamma globulin. A.S. No. 1341736, priority 28.03.85.
3. Schrivener, Art, Lev, Ivekoviceva, PKa, Ackbarali, Justforyou way to obtain the USSR beta1-glycoprotein. A.S. No. 1783644, priority 04.12.89.
4. MeV, Schrivener, Justiniano. Comparative physico-chemical characteristics of drugs USSR β1-glycoprotein isolated from retroplatsentarno blood. J. "Bulletin of experimental biology and medicine, 1990, No. 9, s-267.
5. Wailing, Art, Justiniano. "A method of obtaining USSR sub> 1-glycoprotein". A.S. No. 1836957, priority 25.03.90.
The isolation and purification of USSR beta-1-glycoprotein (TBG) of serum retroplatsentarno blood by fractionation with ammonium sulfate, washing and centrifugation, dissolved sediment, affinity chromatography on sepharose with subsequent elution of the target product and its lyophilization, wherein retroplatsentarno blood centrifuged at 10,000 rpm for one hour, the supernatant add sodium chloride to a final concentration of 0.1-1.0 mol/l) and Triton X-100 to a final concentration of 0.01-0.1%, repeat centrifugation, the resulting serum is saturated with ammonium sulfate to 30%, incubated over night under stirring, centrifuged, the precipitate washed three times 30%ammonium sulfate, suspended in 0.15 M phosphate-saline buffer solution pH 7.0 and 7.5, containing 0.5 M sodium chloride and 0.05%Triton X-100, dialist in the ultrafiltration cell, missing a 10-fold volume of buffer solution, then diluted 5 times and spend ultrafiltration with a limit exceptions 100000 D, the resulting permeate is subjected to a single affinity chromatography on sepharose with immobilizovannymi monoclonal antibodies to TBG, elute ofinnovations TBG in 0.1m glycine-HCl buffer solution pH 2,55, the eluate is immediately neutralized, concentrated, dialist in the saline and lyophilizers.
SUBSTANCE: invention refers to medicine, namely to ophthalmology, and aims at treatment of early cataract. A neutral bath of low-molecular peptide fraction that is prepared of acidic protein fraction of muscle extract of mammal lenses fractionated in 100% ammonium sulphate (Vilenzin) is instilled in conjunctival cavity. Instillations are carried out at 9, 12 and 17 o'clock, 2 drops in both eyes twice every 5 minutes within course 2-6 months every 10 days after 30 days.
EFFECT: method provides partial resorption and prevents progression of cataract.
SUBSTANCE: invention represents an immunostimulating pharmaceutical composition containing endotoxine or its endotoxically active part, fetal haemoglobin, or its γ-chain or α,γ-dimer, and optionally, additive(-s) and optionally, a bulking agent; or containing endotoxine or its endotoxically active part; fetal haemoglobin or its γ-chain or α,γ-dimer; optionally, other compound(-s); and optionally, additive(-s), and a pharmaceutically acceptable carrier and/or a diluent and/or a bulking agent, and containing LPS in amount 10 ng-1 mg and fetal haemoglobin or its partial structures in amount 0.001-10 mg.
EFFECT: synergistic medicobiological activity.
22 cl, 15 ex, 14 tbl, 60 dwg
SUBSTANCE: invention concerns medicine, arthrology. Said therapy includes application of ointment on the joint area. The ointment contain includes Diclofenac, Lydasa, Indometacin, Cinnarizine, Allomaron, Baralgin, a Cerebrolysin, Nootropil, calendula extract, chestnut extract, Dalargin, Solbrol, Extrapone, Eftillinum, sulphidic silt mud in certain quantitative ratio. Ointment is kept for 20-30 minutes. Sessions are daily, once a day. Therapeutic course includes 10-12 procedures.
EFFECT: method improves clinical effectiveness ensured by improved absorbability of sulphide silt mud and other biologically active substances.
SUBSTANCE: way of spinochrome A reception and the sea urchins protein interacting with polyhydroxynaphtoquinone is offered, characterised that raw materials are wash with water, extracted using ethyl alcohol, demineralised with 20% phosphoric acid; then the water-acid admixture is filtered, centrifugated, further the received solution is placed on polychrome 1, admixtures are washed out with 3% phosphoric acid and the distilled water, and the complex of protein with polyhydroxynaphtoquinone is eluted by 10% ethyl alcohol, then 40% ethyl alcohol, fractions are aggregated; further ethyl alcohol is removed, the water residue is freeze dried; received lyophilic powder is dissolved in the water, a non-dissolved deposit is separated, and ethyl alcohol is added to an aqueous solution at a parity 1:9 and maintained within 18-24 hours at 4°C, the dropped out protein is separated, ethyl alcohol is washed out and chromatographed on the sephacryl S-200 and freeze dried; then spirituous solutions is aggregated, concentrated to dryness, the dry rest is dissolved in ethyl alcohol and chromatographed using sephadex LH-20, then the violet fraction of spinochrome A is separated, boiled to dryness, the dry residue is dissolved in acetone and crystallised.
EFFECT: expansion of a spectrum of biologically active substances received from sea urchins.
1 ex, 3 dwg
SUBSTANCE: proposed are 60 kDa human heat shock protein peptides, which are an epitope for T-cells, as well as their peptide derivatives, which are modified on sections for contact with MHC molecules, capable of inducing peripheral tolerance mechanisms, particularly energy mechanisms or mechanisms, mediated by clones of T-cells in patients with rheumatoid arthritis. This invention also relates to pharmaceutical compositions containing the said peptides for treating rheumatoid arthritis.
EFFECT: increased effectiveness of compositions and treatment method.
9 cl, 14 dwg, 18 ex
SUBSTANCE: present invention relates to genetic engineering, more specifically to obtaining anticoagulative protein extracted from nematodes (NAP) and can be used in medicine. To obtain a medicinal preparation based on NAP, methanotrophic yeast host cells are cultured, encoding rNAPc2 or rNAPc2/praline, until attaining the desired cell density. NAP is then extracted from the said yeast host cells through cation-exchange chromatography on an expanding layer. To purify the NAP medicinal preparation, hydrophobic-interaction chromatography is used. NAP is extracted and purified at pH levels below 4.
EFFECT: simple and more efficient method of obtaining anticoagulation proteins from nematodes.
25 cl, 8 dwg, 7 tbl, 6 ex
SUBSTANCE: present invention relates to biotechnology. Description is given of a single-strand T-cell receptor (scTCR), containing an α segment, formed by a sequence of a variable region in a TCR chain, joined with the N end of the extracellular sequence with constant region in the TCR chain, a β segment, formed by a sequence of the variable region of the α TCR chain, joined with the N end of the extracellular sequence with constant region of the β TCR chain, and a linker sequence, joining the C end of the α segment with the N end of the β segment, or vice versa. Extracellular sequences of constant regions of α and β segments are joined by a disulphide bond. Extracellular sequences of constant regions can correspond to constant regions of α and β chains of native TCR, cut-off at their C ends such that, cysteic residues, which form the inter-chain native disulphide bond of the TCR, are excluded, or extracellular sequences of constant regions which are in the α and β segments, can correspond to constant regions of α and β chains of native TCR, in which cysteic residues, which form the native inter-chain disulphide bond, are replaced by another amino acid residue, or there is no uncoupled cysteic residue, which is in the β chain of the native TCR. This invention makes available a new class of alpha/beta analogues of scTCR, in which there is a disulphide bond between residues of a single amino acid, contributing to stability of the bond between the alpha and beta regions of the molecule.
EFFECT: such TCR are suitable for screening or for therapeutic purposes.
3 cl, 14 dwg, 3 ex
FIELD: medicine; cosmetology.
SUBSTANCE: claimed is peptide placenta extract, containing proteins, to obtain which triton X100 and polyglukin are used, extract contains 5.0-35.0 mg/ml of protein, 2.0-5.0 mg/ml of glycoproteins and 0.5-1.0 mg/ml of triton X100. Claimed is method of manufacturing peptide placenta extract, including placenta crushing, extraction of biological material with salt solution, heating, centrifuging, sediment separation and further extract purification by filtering, before crushing preliminarily frozen placenta is melted, poured with phosphate buffer solution, into obtained homogenate triton X100 is added, mixed, stood for 3 hours at 4°C, centrifuged at 15000 g at 4°C, after that activated coal and polyglukin are added to extract, mixed, heated mixing continuously, cooled to room temperature, after which mixing is stopped, obtained product is cooled to +4°C, residue is separated and extract, containing target product is centrifuged before filtering.
EFFECT: extension of application range.
SUBSTANCE: present invention refers to medicine, particularly to cardiology and concerns atherosclerosis treatment. For this purpose effective amount of soluble CTLA4 is introduced.
EFFECT: method enables to block the inflammatory processes underlying pathological changes associated with atherosclerosis, due to binding CTLA4 and B7 molecules and prevented interaction with ligands.
18 cl, 21 tbl, 9 ex, 87 dwg
SUBSTANCE: there is disclosed application of compound antagonist VEGF representing VEGFR1R2-FcΔCl(a) or Flt1D2. Flk1D3. FcΔCl (a) and antiproliferative agent: paclitaxel, docetaxel, or cisplatin, carboplatin, iproplatin for making a medical product for cancer treatment and/or tumour growth reduction or inhibition in an indigent person (versions), relevant method, pharmaceutical composition and a kit for cancer treatment.
EFFECT: overtotal effect of the disclosed combination (97% tumour suppression) observed without any by-effects.
10 cl, 2 dwg, 1 tbl
SUBSTANCE: invention refers to ophthalmology and can be used in surgery of subretinal haemorrhages combined with age-specific macular dystrophy with subretinal neovascular membrane. The technique involves subtotal vitrectomy and removal of posterior hyaloid membrane. Bevacizumab and Ranibizumab are injected intravitreally. It is followed with intravitreal injection of tissue plasminogen activator and expanding gas while a patient is pronated.
EFFECT: method allows deploying blood from under retina, ensuring pathogenetic action on haemorrhage source, preventing proliferative processes in vitreal cavity.
SUBSTANCE: group of inventions concerns biology and medicine. There is disclosed method of biotreatment blood serum production involving electrostimulation of an animal, other than a human, blood drawing of specified animal, serum separation from specified blood and gamma irradiation in a dose 10 to 40 kGy. There is offered blood serum product and pharmaceutical composition containing said blood serum product, and also their application for treatment of various diseases.
EFFECT: invention provides production of biotreatment animal's blood serum applied for treatment of wide range of diseases, including epileptic attacks and apoplexy.
24 cl, 4 ex, 7 tbl, 11 dwg
SUBSTANCE: virus-inactivated blood plasma for introduction to the person, in essence, does not contain the agents inactivating viruses, received from donors from whom 10 % or more belong to population of the non-Caucasoid race individuals, thus plasma is received by mixing of blood or blood plasma of A and B groups, it is unessential AB, without adding blood or blood plasmas of the 0 group, and is characterised that it contains: from five to six parts of blood or a blood plasma of the donors having A group of blood, from four to five parts of blood or a blood plasma of the donors having B group of blood, from zero to one part of blood or a blood plasma of the donors having AB group of blood. The pharmaceutical composition for manufacture of a medical product for treatment of deficiencies of factors of coagulation, thrombocytopenic purpura is offered, and for use in a repeated exchange of great volume of plasma. Application a virus-inactivated blood plasma according to the invention for manufacture of a medical product and the way of a virus-inactivated blood plasma manufacture is offered.
EFFECT: rising of efficiency of treatment.
10 cl, 2 ex
SUBSTANCE: in 1st, 9th, 17th and 24th days of therapy, 5-fluorouracil 750 mg and cisplatin 30 mg incubated with autoblood are introduced intravenously. In 7th, 15th and 23rd days, cisplatin 10 mg incubated with autoplasma, Novocaine 0.5% 20 ml, hydrocortisone 1.0 ml, analginum 50% 2.0 ml, vitamin B12 1.0 ml and gentamycin 1.6 g are introduced into presacral of small pelvis. From 10th to 14th days, 2 hours prior to radiotherapy session, a mixture containing Doxorubicin-Ebewe 5 mg, Viferon ointment 2.5 mg, Vinilin 2 ml, Synthomycin emulsion 2 ml and Dimexide 5% 4 ml is supplied to primary-site tumour through a perforated rubber tube. From 18th to 22th days, a mixture containing Mitomycin C 2.5 mg, Viferon ointment 2.5 mg, Vinilin 2 ml, Synthomycin emulsion 2 ml and Dimexide 5% 4 ml is supplied to primary-site tumour through a perforated rubber tube. After an 14-day recess, the therapeutical actions are repeated once again as specified.
EFFECT: involution of tumour, reduced radiation exposure and toxic by-effects of antitumour therapy.
SUBSTANCE: invention concerns ophthalmology and is intended for treatment of cornea edema and other bullous keratopathy manifestations. Medication includes leukocyte suspension of 1200-1500x103 cells/ml in autological serum, where mononuclears comprise 400-570x103 cells per 1 ml of suspension. Additionally medication includes cytokines, interleukines, b-FGF, TGF-b2 growth factors, endogenic TLR ligands, including heat shock proteins. Leukocyte suspension is obtained out of patient blood and activated by polyA:polyU TLR ligand complex (Poludan). Method of medication obtainment involves addition of polyA:polyU (Poludan) solution to patient blood sample at the ratio of 200-400 units per ml, cell activation at +18-+41 C° for 4-8 hours, centrifugation at 100-400G acceleration. Medication is injected in anterior eye chamber in amount of 0.5-1.0 ml until complete substitution of anterior chamber fluid by medication, by intracorneal method if required, and additionally activated autological cell suspension in autological serum is injected in amount of 1.0-2.0 ml paralimbally under conjunctiva in combination with antibiotic.
EFFECT: direct effect on damaged cornea endothelium, stopped or reduced cornea edema and other manifestations of early bullous keratopathy, increased visual acuity.
3 cl, 2 ex
SUBSTANCE: invention concerns medicine, particularly orthopedics and extracorporal treatment methods, and can be applied in aseptic whirlbone necrosis and Perther disease treatment. Method involves venous blood sampling in amount of 300 ml with further blood separation into erythrocyte mass and plasma. Obtained erythrocyte mass is diluted by 200 ml of 0.9% physiological solution and administered intravenously to patient. 100 ml or patient's plasma is placed in thermostat and incubated for 20 minutes at 37°C. Further 20 mg of vasoprostan medicine is added to incubated plasma, and obtained mix is administered to patient by drop infusion for 1.5-2 hours once a day for 10 days.
EFFECT: enhanced efficiency of the pathology treatment due to vasoprostan binding to plasma proteins and prolonged vasoprostan circulation in blood, resulting in arterial blood circulation improvement, venous stasis blocking in aseptic necrosis zone and significant acceleration of whirlbone acceleration.
2 tbl, 2 ex, 4 dwg
SUBSTANCE: invention concerns medicine, particularly vascular surgery, and can be applied in treatment of critical lower extremity ischemia. Method involves sampling of 400-500 ml of blood from vein. Blood is centrifuged for 15 minutes at 2000 rpm rate. Separated erythrocyte mass is dissolved by 200 ml of 0.9% physiological solution and returned to patient intravenously. 20 mcg of vasaprostan medication is added to 200 ml of plasma. Obtained mix is placed in thermostat and incubated for 20 minutes at 37°C, then injected to patient slowly by intravenous drop infusion for 2-3 hours once per day for 10 days.
EFFECT: elevated peripheral arterial pressure, ischemia grade regress, resulting in larger pain-free walking distance.
1 tbl, 2 ex
SUBSTANCE: invention refers to medicine, namely to oncology, and can be used in oesophageal carcinoma therapy. Substance of the invention consist that prior to radiation therapy, autoblood 400 ml is sampled. Plasma is separated by centrifugation. The first bottle is filled with autoplasma 40 ml and 5-fluorouracil 500 mg, while the second and third bottles are filled with autoplasma 40 ml, and the fourth with residual blood corpuscles and methotrexate 20 mg. The second and third bottles are frozen, and the first and fourth bottles are separately incubated for 40 minutes at 37°C. Then mixed autoblood and methotrexate from the fourth bottle are reinfused droplet-intravenously, mixed autoplasma and 5-fluorouracil from the first bottle are introduced into oesophagus through nasoesophageal probe and followed with the first session of radiation therapy. Intraesophageal autoplasma chemotherapy is carried out one day after the autoplasma bottle is unfrozen, added with 5-fluorouracil 500 mg and incubated in a thermostat. The entire therapy cycle is carried out for the following weeks of radiation therapy; irradiation is involved to TBD (total basic dose) 40-60 Gy at a single dose 2 Gy including 5 sessions a week.
EFFECT: method allows improving effectiveness of radiation therapy ensured by direct antineoplastic exposure of intraesophageal autoplasmochemotherapy and radiosensitisation of autohemochemotherapy.
SUBSTANCE: invention refers to medicine, namely to ceruloplasmin recovery, and can be used in making enzyme blood-plasma medicines. The disclosed method includes dissolution of feed stock in acetate buffer, enzyme sorption in an ion-exchange reactor with functional DEAE group, elution and following purification. Said ion-exchange reactor with functional DEAE group represents a sorbent on fixed styroldivinylbenzene matrix with co-graft DEAE group. And in purification, at first the dissolved enzyme is treated with a solvent-detergent mixture. Then the treated enzyme is immobilised on sulphopropylcation sorbent, washed in acetate buffer containing sodium chloride. The purified enzyme is eluted, while the feed stock is dissolved in acetate solution. The enzyme is sorbed in the ion-exchange reactor wherein as such sorbent on fixed styroldivinylbenzene matrix with co-graft DEAE group is applied. The enzyme is eluted and purified. In purification, the enzyme solution is treated with a solvent-detergent mixture, immobilised on sulphopropylcation sorbent, washed in acetate buffer containing sodium chloride. The purified enzyme is eluted.
EFFECT: method ensures virus safety of recovered medical product ceruloplasmin and allows improving end product yield.
9 cl, 2 ex
SUBSTANCE: invention refers to medicine, namely to virus inactivation in donor enzyme ceruloplasmin production. The disclosed method involves chromatographic purification of enzyme solution by washing with acetate buffer thereafter followed with elution. The enzyme solution is prepared with a solvent-detergent mixture and pre-washed in two stages while being immobilised on sulphopropylcation sorbent. And the first stage includes application of acetate buffer containing sodium caprilate, sodium chloride and propylene glycol, while the second stage involves acetate buffer containing sodium chloride.
EFFECT: invention provides high effectiveness in virus removing and inactivation thereby improving virus safety ceruloplasmin.
9 cl, 6 ex
FIELD: medicine, ophthalmology.
SUBSTANCE: one should apply an autohemocomponent preparation being supernatant liquid of patient's autoblood at increased serotonin content obtained due to irreversible thrombocytic aggregation due to the impact of 0.5 mg ATP per 1.0 ml plasma followed by a 30-min-long centrifuging at the rate of 1000, 2000 and 3000 rot./min for 20, 7 and 3 min, correspondingly. In case of no exudative phenomena on patient's eye bottom the obtained preparation should introduced at the quantity of 7-10 ml once in 48 h for 1 mo (totally, 15 injections). In case of exudative-hemorrhagic phenomena it should be introduced parabulbarly at the volume of 0.5 ml and parenterally - 7.0-10.0 ml once in 48 h for 1 mo per 15 injections, correspondingly. The preparation enables to improve visual functions due to decreased tissue hypoxia and normalization of microcirculation in visual analyzer.
EFFECT: higher efficiency of therapy.
2 cl, 7 dwg, 2 ex